Abstract: Background and Objective: The pathogenesis of osteoarthritis (OA) is highly complex and remains unclear. The pathogenesis of osteoarthritis in an IL-1β-induced human chondrocyte model was studied by transcriptome sequencing in this research. Materials and Methods: An osteoarthritis model was established by the treatment of C28/I2 cells with IL-1β. Using a CCK-8 assay, the concentration of IL-1β at which C28/I2 cells were stimulated was determined to be 10 ng mL–1. Transcriptome sequencing was performed on the cells treated with 10 ng mL–1 IL-1β for 24 hrs. The expression of three randomly screened differentially expressed genes was verified by qRT-PCR. Results: Transcriptome sequencing indicated that the treatment resulted in a total of 5.656 differentially expressed genes, of which 2.891 were up-regulated and 2.765 down-regulated. The results of qRT-PCR were consistent with the results of sequencing. KEGG analysis identified the IL-17 signalling pathway, apoptosis, the NF-kappa B signalling pathway and the Toll-like receptor signalling pathway as being closely related to the pathogenesis of OA. Conclusion: A relationship diagram of differential genes and signalling pathway networks indicated that the pathogenesis of OA may be the result of the interaction and common regulation of differential genes and multiple pathways. This research will provide the theoretical basis for subsequent molecular research on the pathogenesis of OA.